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. 2016 Dec 23;8(1):4. doi: 10.3390/genes8010004

Figure 4.

Figure 4

Knockdown of miR-203 and miR-320b up regulates Dlx5, Runx2 and Osx in osteoblast differentiation. Primary human bone cells were transfected with pre-miRNA-203 and pre-miR-320b and RNA and protein isolated at different time points. q-PCR analysis of: Dlx5 (A); Runx2 (C); and Osx (E) showed up regulation when compared to the pre-miRNA negative control. mRNA levels were normalized to GAPDH. Up regulation at the protein levels of: (B) Dlx5, Lane 1, BMP-2 (+); lane 2, BMP-2 (−); (D) Runx2, Lane 1, BMP-2 (−); lane 2, BMP-2 (+); and (F) Osx, Lane 1, BMP-2 (+); lane 2, BMP-2 (−); lane 3, BMP-2 (+) and anti-miRNA negative control; lane 4, BMP-2 (+) and anti-miR-203; lane 5, BMP-2 (+) and anti-miR-320b; lane 6, BMP-2 (+) and anti-miR-203 and -320b; with GAPDH as a loading control. All values are represented as mean ± S.D. of three independent experiments. Statistical significance is indicated as * p < 0.05 and ** p < 0.001.